Composition for Preventing or Treating Cervical Cancer Having Human Papillomavirus Plasmodium and Immunity Enhancer
a technology of human papillomavirus and plasmodium plasmodium, which is applied in the direction of viruses, drug compositions, peptides, etc., can solve the problems of poor therapeutic effect of composition, and achieve the effect of improving immunogenicity
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example 1
Construction of GX-188E DNA
[0068]Abbreviations used in the Examples of the present invention are defined as follows. An optimized nucleic acid sequence, “tPa” or “t” refers to a secretory signal sequence of a tissue plasminogen activator, and “F” or “Flt3L” refers to an fms-like tyrosine kinase-3 ligand.
[0069]A codon-optimized tPa secretory signal sequence having a nucleic acid sequence set forth in SEQ ID NO: 5 and a codon-optimized Flt3L having a nucleic acid sequence set forth in SEQ ID NO: 6 were chemically synthesized while the codon-optimized tPa and Flt3L secretory signal sequences were coupled to each other. A terminus of the synthesized signal sequence was provided with KpnI (5′) and NheI (3′) restriction sites to facilitate insertion into a vector. A vector for preparing a DNA vaccine, that is, pGX10 (Korean Patent Publication No. 2003-0047667) was digested with KpnI and NheI restriction enzymes, and then ligated with the synthesized tPa-Flt3L signal sequence using ligase ...
example 2
Confirmation of Therapeutic Effect of GX-188E on Cervical Cancer
[0071]To confirm a therapeutic effect of GX-188E on cervical cancer, TC-1 tumor cells were subcutaneously injected into C57BL / 6 rats at a concentration of 5×105 cells, and GX-188E was intramuscularly injected at doses of 50 μg and 100 μg on days 3 and 8, followed by performing electroporation. A change in volume of tumor cells was observed from the day of injection to day 27, and the spleens of the rats were extracted on Day 36. Then, 1×106 cells were put into a plate coated with 50 μl of a 5 μg / ml anti-mouse IFN-g antibody (BD Pharmigen, San Diego, Calif.) together with IL-2 and an HPV16 E6 CD8 T cell epitope (E648-57; EVYDFAFRDL, Peptron, Korea), an HPV18 E7 CD8 T cell epitope (E749-57; RAHYNIVTF, Peptron, Korea), an HPV18 E6 peptide pool, or an HPV18 E7 peptide pool, and incubated at 37° C. for 24 hours in a 5% CO2 incubator (Froma, Minn., USA). The plate was washed with PBST, and a 2 μg / ml IFN-g detection antibody c...
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